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体外雌二醇-受体复合物对克隆的禽卵黄生成素II基因转录的特异性调控

Specific modulation of the transcription of cloned avian vitellogenin II gene by estradiol-receptor complex in vitro.

作者信息

Jost J P, Geiser M, Seldran M

出版信息

Proc Natl Acad Sci U S A. 1985 Feb;82(4):988-91. doi: 10.1073/pnas.82.4.988.

Abstract

Avian vitellogenin-cauliflower mosaic virus hybrid gene is effectively transcribed in vitro in the homologous embryonic liver nuclei system. The transcription of the hybrid gene is modulated by the addition of an estradiol-receptor preparation that has been shown to bind selectively to an upstream region of cloned vitellogenin gene. Stimulation of the transcription of cloned vitellogenin hybrid gene by estradiol receptor is alpha-amanitin sensitive, hormone dependent, and promoter specific. Simian virus 40 and Escherichia coli promoters are not stimulated by the estradiol-receptor complex. The endogenous silent vitellogenin II gene (wild type) present in the nuclei is not turned on by the addition of estradiol-receptor complex. Deletion or inversion of the DNA sequence where the estradiol-receptor complex binds results in the complete suppression of the in vitro stimulation of transcription by estradiol receptor. Correct initiation of the transcription was demonstrated by primer extension studies of the newly synthesized RNA.

摘要

禽卵黄生成素-花椰菜花叶病毒杂交基因在同源胚胎肝细胞核系统中能有效地进行体外转录。杂交基因的转录可通过添加一种雌二醇受体制剂来调节,该制剂已被证明能选择性地结合克隆的卵黄生成素基因的上游区域。雌二醇受体对克隆的卵黄生成素杂交基因转录的刺激是α-鹅膏蕈碱敏感的、激素依赖性的且具有启动子特异性。猿猴病毒40和大肠杆菌启动子不会被雌二醇-受体复合物刺激。细胞核中存在的内源性沉默卵黄生成素II基因(野生型)不会因添加雌二醇-受体复合物而被激活。雌二醇-受体复合物结合的DNA序列的缺失或倒置会导致雌二醇受体对转录的体外刺激完全受到抑制。通过对新合成RNA的引物延伸研究证明了转录的正确起始。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ffa/397178/d9bb282d2c89/pnas00344-0043-a.jpg

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